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Photographs showing a comparison of the drought resistance state of the transgenic and WT plants. (A) Effects of drought on the transgenic and WT plants. (B) The survival rate of the transgenic and WT plants after being rewatered for 7 days. (C) The proline content. (D) The MDA content. Error bars represent ±SD, and each value represents the averages of three repeated measurements. ** indicate significant differences between the transgenic plants and the WT plants (p < 0.01).
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bZIP transcription factors have been reported to be involved in many different biological processes in plants. The ABA (abscisic acid)-dependent AREB/ABF-SnRK2 pathway has been shown to play a key role in the response to osmotic stress in model plants. In this study, a novel bZIP gene, FtbZIP5, was isolated from tartary buckwheat, and its role in t...
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Context 1
... confirm the function of FtbZIP5 in Arabidopsis stress resistance, the phenotypic and physiological changes in the transgenic lines and the WT were analyzed after drought, salt and oxidative stress. The initial number of plants used before treatment was 30 plants per pot, and the survival rate of transgenic lines (L2, L4 and L6) after 3 weeks of dewatering and 7 days of rewatering was approximately 60% higher than that of WT ( Figure 5B). In addition, the survival rate of the WT plants was at least 25% lower than that of the L2, L4 and L6 plants after 3 weeks of salt stress treatment. ...
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... addition, the survival rate of the WT plants was at least 25% lower than that of the L2, L4 and L6 plants after 3 weeks of salt stress treatment. Furthermore, the levels of the antioxidant enzymes (SOD, POD and CAT) and the proline content in the two-week-old transgenic subjected to drought conditions or treated with salt were more insensitive than those of the WT plants ( Figure 5C, Figure 6C, Figure 7C-E), while the content of MDA was more insensitive than that of the WT ( Figure 5D, Figure 6D). Consistently, the results showed that the chlorophyll degradation rate in the transgenic lines was lower than that of the WT at MV concentration of 10 µM and 30 µM MV, as shown in Figure 8A,B. ...
Context 3
... addition, the survival rate of the WT plants was at least 25% lower than that of the L2, L4 and L6 plants after 3 weeks of salt stress treatment. Furthermore, the levels of the antioxidant enzymes (SOD, POD and CAT) and the proline content in the two-week-old transgenic subjected to drought conditions or treated with salt were more insensitive than those of the WT plants ( Figure 5C, Figure 6C, Figure 7C-E), while the content of MDA was more insensitive than that of the WT ( Figure 5D, Figure 6D). Consistently, the results showed that the chlorophyll degradation rate in the transgenic lines was lower than that of the WT at MV concentration of 10 µM and 30 µM MV, as shown in Figure 8A,B. ...
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Citations
... An increasing number of studies have revealed that the SnRK family is a crucial component in plant signaling transduction and response to abiotic stress [50,51]. Moreover, the SnRK family members of many plants have been clarified, including Liriodendron chinense [52], Brassica napus [53], Casuarina equisetifolia [54], Brachypodium distachyon [55], cotton [56], Triticum aestivum [57] and Phaseolus vulgaris [58]. ...
The SnRK gene family is the chief component of plant stress resistance and metabolism through activating the phosphorylation of downstream proteins. S. miltiorrhiza is widely used for the treatment of cardiovascular diseases in Asian countries. However, information about the SnRK gene family of S. miltiorrhiza is not clear. The aim of this study is to comprehensively analyze the SnRK gene family of S. miltiorrhiza and its response to phytohormone. Here, 33 SmSnRK genes were identified and divided into three subfamilies (SmSnRK1, SmSnRK2 and SmSnRK3) according to phylogenetic analysis and domain. SmSnRK genes within same subgroup shared similar protein motif composition and were unevenly distributed on eight chromosomes of S. miltiorrhiza. Cis-acting element analysis showed that the promoter of SmSnRK genes was enriched with ABRE motifs. Expression pattern analysis revealed that SmSnRK genes were preferentially expressed in leaves and roots. Most SmSnRK genes were induced by ABA and MeJA treatment. Correlation analysis showed that SmSnRK3.15 and SmSnRK3.18 might positively regulate tanshinone biosynthesis; SmSnRK3.10 and SmSnRK3.12 might positively regulate salvianolic acid biosynthesis. RNAi-based silencing of SmSnRK2.6 down-regulated the biosynthesis of tanshinones and biosynthetic genes expression. An in vitro phosphorylation assay verified that SmSnRK2.2 interacted with and phosphorylated SmAREB1. These findings will provide a valuable basis for the functional characterization of SmSnRK genes and quality improvement of S. miltiorrhiza.
... Furthermore, Salicylic acid (SA) is an important plant hormone that participates in the plant's response to stress through complex signal transduction networks [74]. Plant bZIP-type transcription factors are induced in their expression by SA [75]. ...
Salt stress is a significant environmental factor affecting plant growth and development, with NaCl stress being one of the most common types of salt stress. The halophyte, Tamarix ramosissima Ledeb (T. ramosissima), is frequently utilized for the afforestation of saline-alkali soils. Indeed, there has been limited research and reports by experts and scholars on the regulatory mechanisms of basic leucine zipper (bZIP) genes in T. ramosissima when treated with exogenous potassium (K+) to alleviate the effects of NaCl stress. This study focused on the bZIP genes in T. ramosissima roots under NaCl stress with additional KCl applied. We identified key candidate genes and metabolic pathways related to bZIP and validated them through quantitative real-time PCR (qRT-PCR). The results revealed that under NaCl stress with additional KCl applied treatments at 0 h, 48 h, and 168 h, based on Pfam protein domain prediction and physicochemical property analysis, we identified 20 related bZIP genes. Notably, four bZIP genes (bZIP_2, bZIP_6, bZIP_16, and bZIP_18) were labeled with the plant hormone signal transduction pathway, showing a predominant up-regulation in expression levels. The results suggest that these genes may mediate multiple physiological pathways under NaCl stress with additional KCl applied at 48 h and 168 h, enhancing signal transduction, reducing the accumulation of ROS, and decreasing oxidative damage, thereby enhancing the tolerance of T. ramosissima to NaCl stress. This study provides gene resources and a theoretical basis for further breeding of salt-tolerant Tamarix species and the involvement of bZIP transcription factors in mitigating NaCl toxicity.
... In transgenic tobacco overexpressing SoSnRK2.1, under drought stress, ionic liquid (IL), MDA, hydrogen peroxide (H 2 O 2 ), superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities increased [11], indicating that overexpression of SoSnRK2.1 enhanced the drought tolerance of tobacco. Another study found that the Fagopyrum tataricum transcription factor FtbZIP5 was regulated by FtSnRK2.6 to enhance drought tolerance in transgenic Arabidopsis [12]. Katsuta et al. [13] observed that the abiotic stress response of RAF kinase plays a critical role in the SnRK2-mediated osmotic stress response by interacting with AtSnRK2.6, a central factor in stomatal closure under drought conditions, to protect plants from drought injury. ...
Drought stress restricts vegetable growth, and abscisic acid plays an important role in its regulation. Sucrose non-fermenting1-related protein kinase 2 (SnRK2) is a key enzyme in regulating ABA signal transduction in plants, and it plays a significant role in response to multiple abiotic stresses. Our previous experiments demonstrated that the SnRK2.11 gene exhibits a significant response to drought stress in cucumbers. To further investigate the function of SnRK2.11 under drought stress, we used VIGS (virus-induced gene silencing) technology to silence this gene and conducted RNA-seq analysis. The SnRK2.11-silencing plants displayed increased sensitivity to drought stress, which led to stunted growth and increased wilting speed. Moreover, various physiological parameters related to photosynthesis, chlorophyll fluorescence, leaf water content, chlorophyll content, and antioxidant enzyme activity were significantly reduced. The intercellular CO2 concentration, non-photochemical burst coefficient, and malondialdehyde and proline content were significantly increased. RNA-seq analysis identified 534 differentially expressed genes (DEGs): 311 were upregulated and 223 were downregulated. GO functional annotation analysis indicated that these DEGs were significantly enriched for molecular functions related to host cells, enzyme activity, and stress responses. KEGG pathway enrichment analysis further revealed that these DEGs were significantly enriched in phytohormone signalling, MAPK signalling, and carotenoid biosynthesis pathways, all of which were associated with abscisic acid. This study used VIGS technology and transcriptome data to investigate the role of CsSnRK2.11 under drought stress, offering valuable insights into the mechanism of the SnRK2 gene in enhancing drought resistance in cucumbers.
... FlbZIP12 shares high sequence similarity to the Tartary buckwheat transcription factor FtbZIP5 ( Figure S1). In a previous study, FtbZIP5 was shown to interact with FtSnRK2.6 in yeast (S. cerevisiae) (Li et al., 2020). Thus, we hypothesized that FlbZIP12 might interact with FlSnRK2s, including FlSnRK2.2, FlSnRK2.3, and FlSnRK2.6. ...
Karst lands provide a poor substrate to support plant growth, as they are low in nutrients and water content. Common buckwheat (Fagopyrum esculentum) is becoming a popular crop for its gluten-free grains and their high levels of phenolic compounds, but buckwheat yields are affected by high water requirements during grain filling. Here, we describe a wild population of drought-tolerant Fagopyrum leptopodum and its potential for enhancing drought tolerance in cultivated buckwheat. We determined that the expression of a gene encoding a Basic leucine zipper (bZIP) transcription factor, FlbZIP12, from F. leptopodum is induced by abiotic stresses, including treatment with the phytohormone abscisic acid, salt, and polyethylene glycol. In addition, we show that overexpressing FlbZIP12 in Tartary buckwheat (Fagopyrum tataricum) root hairs promoted drought tolerance by increasing the activities of the enzymes superoxide dismutase and catalase, decreasing malondialdehyde content, and upregulating the expression of stress-related genes. Notably, FlbZIP12 overexpression induced the expression of key genes involved in flavonoid biosynthesis. We also determined that FlbZIP12 interacts with protein kinases from the FlSnRK2 family in vitro and in vivo. Taken together, our results provide a theoretical basis for improving drought tolerance in buckwheat via modulating the expression of FlbZIP12 and flavonoid contents.
... F. tataricum is popular as a significant medicine and health food due to its rich content of bioactive flavonoids, which are detected to be highly beneficial for human health [36]. In recent years, studies have characterized a set of modulators playing crucial roles in the growth and development of F. tataricum under certain environmental factors [37][38][39][40][41][42][43]. For example, FtMYB6 and FtMYB116, were found to be positive regulators of flavonoids biosynthesis in the light [37,38]. ...
... For example, FtMYB6 and FtMYB116, were found to be positive regulators of flavonoids biosynthesis in the light [37,38]. The FtMYB9, FtMYB13, FtbZIP5, and FtbZIP83 were identified as positive regulators of drought and salt response and improved drought and salt tolerance of transgenic Cruciformes Cruciferae Arabidopsis thaliana (L.) Heynh., while FtMYB10 was found to be a negative regulator of drought and salt response in transgenic Arabidopsis thaliana [39][40][41][42][43]. However, previous studies mainly focused on the transcriptional and post-transcriptional levels [37][38][39][40][41][42][43], and thus the knowledge of the pre-transcriptional level (epigenetic level) is extremely limited. ...
... The FtMYB9, FtMYB13, FtbZIP5, and FtbZIP83 were identified as positive regulators of drought and salt response and improved drought and salt tolerance of transgenic Cruciformes Cruciferae Arabidopsis thaliana (L.) Heynh., while FtMYB10 was found to be a negative regulator of drought and salt response in transgenic Arabidopsis thaliana [39][40][41][42][43]. However, previous studies mainly focused on the transcriptional and post-transcriptional levels [37][38][39][40][41][42][43], and thus the knowledge of the pre-transcriptional level (epigenetic level) is extremely limited. ...
Histone deacetylases (HDACs), known as histone acetylation erasers, function crucially in plant growth and development. Although there are abundant reports focusing on HDACs of Arabidopsis and illustrating their important roles, the knowledge of HDAC genes in Tartary buckwheat (Polygonales Polygonaceae Fagopyrum tataricum (L.) Gaertn) is still scarce. In the study, a total of 14 HDAC genes were identified and divided into three main groups: Reduced Potassium Dependency-3/His-52 tone Deacetylase 1 (RPD3/HDA1), Silent Information Regulator 2 (SIR2), and the plant-53 specific HD2. Domain and motif composition analysis showed there were conserved domains and motifs in members from the same subfamilies. The 14 FtHDACs were distributed asymmetrically on 7 chromosomes, with three segmental events and one tandem duplication event identified. The prediction of the cis-element in promoters suggested that FtHDACs probably acted in numerous biological processes including plant growth, development, and response to environmental signals. Furthermore, expression analysis based on RNA-seq data displayed that all FtHDAC genes were universally and distinctly expressed in diverse tissues and fruit development stages. In addition, we found divergent alterations in FtHDACs transcript abundance in response to different light conditions according to RNA-seq and RT-qPCR data, indicating that five FtHDACs might be involved in light response. Our findings could provide fundamental information for the HDAC gene family and supply several targets for future function analysis of FtHDACs related with light response of Tartary buckwheat.
... In addition, FtZIP5 interacts with Ft-SnRK2.6 [32]. In this study, we found that FtZIP85 was notably expressed in the seeds ( Figure 1). ...
... In addition, there are few studies on the influence of the flavonoid synthesis pathway substances in Tartary buckwheat acting via interactions with the important response genes of the ABA pathway. Only two bZIP TFs have been preliminarily noted to interact with SnRK2s [22,32]. In this study, we reported a novel bZIP TF, FtbZIP85, that positively regulates PA biosynthesis. ...
As a drought-tolerant crop, Tartary buckwheat survives under adverse environmental conditions, including drought stress. Proanthocyanidins (PAs) and anthocyanins are flavonoid compounds, and they participate in the regulation of resistance to both biotic and abiotic stresses by triggering genes’ biosynthesis of flavonoids. In this study, a basic leucine zipper, basic leucine zipper 85 (FtbZIP85), which was predominantly expressed in seeds, was isolated from Tartary buckwheat. Our study shows that the expressions of FtDFR, FtbZIP85 and FtSnRK2.6 were tissue-specific and located in both the nucleus and the cytosol. FtbZIP85 could positively regulate PA biosynthesis by binding to the ABA-responsive element (ABRE) in the promoter of dihydroflavonol 4-reductase (FtDFR), which is a key enzyme in the phenylpropanoid biosynthetic pathway. Additionally, FtbZIP85 was also involved in the regulation of PA biosynthesis via interactions with FtSnRK2.6 but not with FtSnRK2.2/2.3. This study reveals that FtbZIP85 is a positive regulator of PA biosynthesis in TB.
... The rest of the plants were watered with 200 mM NaCl for 7 and 12 days. Meanwhile, the survival rate, water loss rate, fresh weight, proline (Pro), malondialdehyde (MDA), ion leakage (IL), superoxide anion (O 2 − ), hydrogen peroxide (H 2 O 2 ), diaminobenzidine (DAB) and nitroblue tetrazolium (NBT) staining were measured in the transgenic Arabidopsis plants and WTs after all the processing was completed as previously described [51]. To investigate whether stomatal pore closure was sensitive to ABA, the stomatal aperture was visualized according to the methods described previously [52,53]. ...
Tartary buckwheat (Fagopyrum tataricum Gaertn.) is a coarse cereal with strongly abiotic resistance. The MYB family plays a regulatory role in plant growth, development, and responses to biotic and abiotic stresses. However, the characteristics and regulatory mechanisms of MYB transcription factors in Tartary buckwheat remain unclarified. Here, this study cloned the FtMYB22 gene from Tartary buckwheat, and investigated its involvement in responding to individual water deficit and salt stress in Arabidopsis. Sequence analysis highlighted that the N-termini of FtMYB22 contained two highly conserved SANT domains and one conserved domain from the SG20 subfamily. Nucleus-localized FtMYB22 did not have individual transcriptional activation activity. Water deficiency and salt stress induced the high expression of the GUS gene, which was driven by the promoter of FtMYB22. Yeast stress experiments showed that the overexpression of FtMYB22 significantly reduced the growth activity of transgenic yeast under water deficit or salt stress. Consistently, the overexpression of FtMYB22 reduced the salt and water deficit stress resistance of the transgenic plants. In addition, physiological parameters showed that transgenic plants had lower proline and antioxidant enzyme activity under stress conditions. Compared to the wild-type (WT), transgenic plants accumulated more malondialdehyde (MDA), H2O2, and O2−; they also showed higher ion permeability and water loss rates of detached leaves under stress treatments. Notably, FtMYB22 was involved in plant stress resistance through an ABA-dependent pathway. Under stress conditions, the expression of RD29A, RD29B, PP2CA, KIN1, COR15A, and other genes in response to plant stress in transgenic lines was significantly lower than that in the WT (p < 0.05). Furthermore, yeast two-hybrid assay showed that there was a significant interaction between FtMYB22 and the ABA receptor protein RCAR1/2, which functioned in the ABA signal pathway. Altogether, FtMYB22, as a negative regulator, inhibited a variety of physiological and biochemical reactions, affected gene expression and stomatal closure in transgenic plants through the ABA-dependent pathway, and reduced the tolerance of transgenic Arabidopsis to water deficiency and salt stress. Based on these fundamental verifications, further studies would shed light on the hormone signal response mechanism of FtMYB22.
... The expression of FtbZIP12 was induced by ABA, NaCl, and mannitol; however, the RT-qPCR results showed that changes in the FtbZIP12 expression after 0-24 h of mannitol induction were different from those under NaCl stress (Figure 2A,B). This expression pattern was different from that of FtbZIP83 and FtbZIP5, the relative expression of which first increased then decreased under ABA, NaCl, and drought stress [26,27]. We concluded that although FtbZIP12 belongs to the same subfamily as FtbZIP83 and FtbZIP5, they are not only different in sequence, but also have different roles under osmotic stress. ...
ABFs play a key role in regulating plant osmotic stress. However, in Tartary buckwheat, data on the role of ABF genes in osmotic stress remain limited and its associated mechanism in osmoregulation remain nebulous. Herein, a novel ABF family in Tartary buckwheat, FtbZIP12, was cloned and characterized. FtbZIP12 is a transcriptional activator located in the nucleus; its expression is induced by NaCl, mannitol, and abscisic acid (ABA). Atopic expression of FtbZIP12 in Arabidopsis promoted seed germination, reduced damage to primary roots, and improved the tolerance of seedlings to osmotic stress. The quantitative realtime polymerase chain reaction (RT-qPCR) results showed that the expressions of the typical genes related to stress, the SOS pathway, and the proline synthesis pathway in Arabidopsis were significantly (p < 0.05) upregulated under osmotic stress. FtbZIP12 improved the osmotic pressure resistance by reducing the damage caused by reactive oxygen species to plants and maintained plant homeostasis by upregulating the expression of genes related to stress, osmotic regulation, and ion homeostasis. This study identified a key candidate gene for understanding the mechanism underlying osmotic-stress-regulated function in Tartary buckwheat, thereby providing a theoretical basis for improving its yield and quality.
... Among these TFs, two bZIP TF genes (FtPinG0003196200.01/FtbZIP5 and FtPinG0002143600.01/ FtbZIP83), which are homologous to AtABF2, have been demonstrated to play a positive regulatory role in drought resistance of transgenic Arabidopsis (Li et al., 2019b;Li et al., 2020). Four NAC TF genes (FtPinG0002561000.01, ...
... Notably, among these TFs, two bZIP TFs (FtPinG0003196200.01/FtbZIP5 and FtPinG0002143600.01/FtbZIP83) have been demonstrated to play a positive regulatory role in drought resistance in transgenic Arabidopsis (Li et al., 2019b;Li et al., 2020). Four NAC TFs are the homologs of Arabidopsis drought-resistant TFs (ANAC019, ANAC72/RD26, NAC029, and ANAC2, respectively) (Fujita et al., 2011). ...
Drought stress is one of the major abiotic stress factors that affect plant growth and crop productivity. Tartary buckwheat is a nutritionally balanced and flavonoid-rich pseudocereal crop and also has strong adaptability to different adverse environments including drought. However, little is known about its drought tolerance mechanism. In this study, we performed comparative physiological and transcriptomic analyses of two contrasting drought-resistant Tartary buckwheat genotypes under nature drought treatment in the reproductive stage. Under drought stress, the drought-tolerant genotype XZSN had significantly higher contents of relative water, proline, and soluble sugar, as well as lower relative electrolyte leakage in the leaves than the drought-susceptible LK3. A total of 5,058 (2,165 upregulated and 2,893 downregulated) and 5,182 (2,358 upregulated and 2,824 downregulated) potential drought-responsive genes were identified in XZSN and LK3 by transcriptome sequencing analysis, respectively. Among the potential drought-responsive genes of XZSN, 1,206 and 1,274 genes were identified to be potential positive and negative contributors for XZSN having higher drought resistance ability than LK3. Furthermore, 851 out of 1,206 positive drought-resistant genes were further identified to be the core drought-resistant genes of XZSN based on WGCNA analysis, and most of them were induced earlier and quicker by drought stress than those in LK3. Functional annotation of the 851 core drought-resistant genes found that a large number of stress-responsive genes were involved in TFs, abscisic acid (ABA) biosynthesis, signal transduction and response, non-ABA signal molecule biosynthesis, water holding, oxygen species scavenging, osmotic adjustment, cell damage prevention, and so on. Transcriptional regulatory network analyses identified the potential regulators of these drought-resistant functional genes and found that the HD-ZIP and MYB TFs might be the key downstream TFs of drought resistance in Tartary buckwheat. Taken together, these results indicated that the XZSN genotype was more drought-tolerant than the LK3 genotype as evidenced by triggering the rapid and dramatic transcriptional reprogramming of drought-resistant genes to reduce water loss, prevent cell damage, and so on. This research expands our current understanding of the drought tolerance mechanisms of Tartary buckwheat and provides important information for its further drought resistance research and variety breeding.
... The AREB/ABF transcription factors not only regulate plant growth and development, but also play an important role in plant response to abiotic stresses such as drought, low temperature and high salt concentration. Previous studies showed that AtABFs could promote ABA-mediated chlorophyll [20][21][22]degradation and leaf senescence [23]. Overexpressing NtABF induced the up-regulation of reactive oxygen species (ROS) scavenging gene and improved the antioxidant ability of tobacco [24]. ...
Background
Plants suffer from various abiotic stresses during their lifetime, of which drought and salt stresses are two main factors limiting crop yield and quality. Previous studies have shown that abscisic acid (ABA) responsive element binding protein (AREB)/ ABRE binding factors (ABFs) in bZIP transcription factors are involved in plant stress response in an ABA-dependent manner. However, little is known about the properties and functions of AREB/ABFs, especially ABF3, in cotton.
Results
Here, we reported the cloning and characterization of GhABF3. Expression of GhABF3 was induced by drought,salt and ABA treatments. Silencing of GhABF3 sensitized cotton to drought and salt stress, which was manifested in decreased cellular antioxidant capacity and chlorophyll content. Overexpression of GhABF3 significantly improved the drought and salinity tolerance of Arabidopsis and cotton. Exogenous expression of GhABF3 resulted in longer root length and less leaf wilting under stress conditions in Arabidopsis thaliana. Overexpressing GhABF3 significantly improved salt tolerance of upland cotton by reducing the degree of cellular oxidation, and enhanced drought tolerance by decreasing leaf water loss rate. The increased expression of GhABF3 up-regulated the transcriptional abundance of downstream ABA-inducible genes under salt stress in Arabidopsis.
Conclusion
In conclusion, our results demonstrated that GhABF3 plays an important role in plant drought and salt tolerance. Manipulation of GhABF3 by biotechnology might be an important strategy to alter the stress resistance of cotton.
